SBIR Phase I: Development of a Novel Biocontainment/Biosafety Platform Using Synthetic Auxotrophs

这个小企业创新研究（SBIR）第一阶段项目的更广泛的影响是开发一个新的系统，将提高转基因生物（GMOs）的安全性和生产力。转基因生物使新的活性治疗剂、农产品和生物基消费品成为可能。然而，一个关键的挑战是缺乏可用的工具来轻松控制转基因生物的行为。该提案将开发一种技术，通过将转基因生物的生存与特定控制化学品的存在联系起来，确保对转基因生物的遏制。核心技术是基于细胞，这些细胞被设计成能够响应特定的化学信号而改变其行为，该项目的目标是扩大可以通过化学输入控制的蛋白质组合（从而改变行为）。这解决了人们对转基因生物的使用和可能逃逸的关切，转基因生物可能产生未知的后果。拟议的项目将建立一个化学控制的必需基因组合。这将通过蛋白质工程和同时构建跨越200个不同必需基因的12，000个蛋白质文库来实现。使用靶向基因组整合，该文库将被整合到大肠杆菌的基因组中。使用阳性和阴性（化学）选择进行文库的表型表征，然后进行下一代DNA测序，将用于揭示单个蛋白质设计的功能（致死性、可行性或条件性）。该数据集将阐明化学控制蛋白质设计的结构、序列和功能之间的关系。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact of this Small Business Innovation Research (SBIR) Phase I project is the development of a novel system that will increase the safety and productivity of genetically modified organisms (GMOs). GMOs enablie novel live therapeutics, agricultural products, and bio-based consumer products. However, a key challenge is the lack of available tools to easily control the behavior of GMOs. This proposal will develop a technology that can be used to ensure containment of GMOs by tying their survival to the presence of specific control chemicals. The core technology is based on cells that have been engineered to alter their behavior in response to specific chemical signals, and the goal of this project is to expand the portfolio of proteins (and thus behaviors) that can be controlled by chemical inputs. This addresses concern surrounding the use and possible escape of GMOs that may have unknown consequences. The proposed project will establish a portfolio of chemically-controlled essential genes. This will be accomplished by protein engineering and simultaneously constructing 12,000 protein libraries spanning 200 different essential genes. Using targeted genome integration, this library of libraries will be integrated into the genome of Escherichia coli. Phenotypic characterization of the library using positive and negative (chemical) selections followed by next-generation DNA sequencing will be used to uncover the function (lethal, viable, or conditional) of individual protein designs. This dataset will illuminate the relationship between structure, sequence, and function for design of chemically-controlled proteins.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.